Preparation and characterization of a monoclonal antibody against HIV-1 inhibitor N-SRCR / 医学研究生学报

ABSTRACT

Objective Protein N-SRCR derived from salivary agglutinin (SAG) inhibits HIV-1 infection.An N-SRCR monoclonal antibody was prepared for the study of the interaction between N-SRCR and HIV-1 envelop glycoprotein (gp120).Methods The purified recombinant N-SRCR expressed by 293 cells was used to immunize four weeks old BALB/c mice.After the final boost,the mouse spleen cells were isolated and fused with mouse myeloma cell line SP2/0-Ag-14,and the resulting hybridomas were screened for the production of N-SRCR-specific antibodies by ELISA assay.The monoclonal antibody against N-SRCR was purified by HiTrap Protein G kit,the purity determined by SDS-PAGE and the antibody titers by ELISA.The antibody specificity.was charqacterized by western blotting.Results A strain of hybridoma cell clones stably secreting N-SRCR antibody,named 1D6,was obtained.The high purity of the IgG was demonstrated by SDS-PAGE,and the ELISA titers of 1D6 was more than 100?25.Conclusion A monoclonal antibody against N-SRCR was successfully prepared,which laid the ground for further studies on the biological function of N-SRCR and the interactions between SRCR domains and HIV-1 Env gp120.